Towards a Mechanistic Undestanding of Adolscent Idiopathic Scoliosis

对青少年特发性脊柱侧凸的机制的理解

• 批准号: 9364384
• 负责人: Ryan Scott Gray
• 金额: $ 34.43万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-10 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9364384
• 关键词: Adolescence; Adolescent; Adult; Affect; Animal Disease Models; Animal Model; Animals; Biological; Biological Models; Bone Tissue; Cartilage; Cell Culture Techniques; Cell Lineage; Cells; Child; Childhood; Collection; Defect; Deformity; Deposition; Development; Disease; Distress; Embryo; Ethylnitrosourea; Etiology; Extracellular Matrix; Fibrosis; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Predisposition to Disease; Grant; Growth and Development function; Homeostasis; Human; Idiopathic scoliosis; Intervertebral disc structure; Left; Life; LoxP-flanked allele; Lung; Massive Parallel Sequencing; Methods; Modeling; Modernization; Molecular; Molecular Genetics; Mouse Strains; Mus; Musculoskeletal Diseases; Mutagenesis; Mutation; Names; Pain; Pathogenesis; Pathway interactions; Population Study; Reagent; Regulation; Research; Signal Transduction; Stem cells; Structure; Testing; Time; Tissue Model; Tissues; Transgenic Organisms; Validation; Vertebral column; Zebrafish; bone cell; cartilage cell; cartilage development; experimental study; forward genetics; gene discovery; genetic approach; genetic resource; genome editing; human population study; insight; mouse model; mutant; novel; postnatal; protein protein interaction; psychosocial; reverse genetics; risk variant; scoliosis

Project Summary: Here we seek to understand the cellular and molecular causes of a common pediatric musculoskeletal disease, adolescent idiopathic scoliosis (AIS). Human population studies of AIS are beginning to uncover important risk loci for this disease, but there has been limited progress in understanding the etiology of AIS. In part, this is due to a lack of good, genetically tractable animal models of scoliotic diseases. We will focus our efforts on the functional analysis of a few explicit animal models of AIS which we generated in both mouse and zebrafish model systems. At the same time, we will advance gene discovery in this field, by applying modern genetic approaches to characterize novel mutations that cause AIS-like spine deformity in an existing collection of zebrafish mutants we generated. The project will test the following hypotheses: First, that late-onset scoliosis in our Gpr126 mouse model is a consequence of embryonic defects of the intervertebral discs. Second, that functional analysis of Gpr126 signaling in cartilage and the intervertebral disc will provide a mechanistic understanding of the pathophysiology of AIS in humans, as well as, help to emphasize new genes and pathways that contribute to AIS. Third, that our plan for gene discovery in the zebrafish model and functional analysis of AIS-like scoliosis in zebrafish and mouse models will provide a synergistic framework for more mechanistic understanding of the causes of AIS in humans. These hypotheses will be tested under three Specific Aims: I. Determine when and where Gpr126 is required for AIS. Loss of Gpr126 function in a common progenitor cell of cartilage and bone tissues models AIS in the mouse. Using conditional genetic approaches in the mouse, we will refine how the loss of Gpr126 in these tissues contribute to the onset and progression of scoliosis and we will define the temporal window for Gpr126 function in spine development. II. What is the molecular function of Gpr126 in the intervertebral disc and cartilage? Here we will investigate the mechanism by which Gpr126 controls the maturation and deposition of the cartialge extracellular matrix and the development of the interverterbal disc and how this relates to onset of scoliosis in the mouse. III. Characterization of the molecular genetics and etiologies of spine deformity. To identify genes and pathways important for normal spine development, we utilized a forward genetics approach to isolate a collection of adult-viable spine deformity mutant zebrafish. We will apply massively parallel sequencing to identify novel spine deformity disease genes in these existing mutant lines. In order to gain a mechanistic understanding of scoliosis we will: a) undertake functional analysis of two of these novel mutant zebrafish lines, affecting extracellular matrix modifying genes; and b) investigate the etiology of scoliosis in an explicit zebrafish model of a well-known human AIS risk locus. The results are excepted to reveal previously unknown mechanisms and pathways essential for normal spine development.

项目摘要：在这里，我们试图了解常见儿科疾病的细胞和分子原因 肌肉骨骼疾病、青少年特发性脊柱侧凸（AIS）。 AIS 的人群研究已经开始 揭示这种疾病的重要风险位点，但在了解病因方面进展有限 AIS 的。部分原因是缺乏良好的、基因上可控制的脊柱侧弯疾病动物模型。我们将 我们的重点是对一些明确的 AIS 动物模型进行功能分析，这些模型是我们在 小鼠和斑马鱼模型系统。同时，我们将推进该领域的基因发现，通过 应用现代遗传学方法来表征导致 AIS 样脊柱畸形的新突变 我们现有的斑马鱼突变体集合。该项目将测试以下假设：首先， 我们的 Gpr126 小鼠模型中的晚发性脊柱侧凸是椎间盘胚胎缺陷的结果 光盘。其次，对软骨和椎间盘中 Gpr126 信号传导的功能分析将提供 对人类 AIS 病理生理学的机制理解，以及有助于强调新基因 以及有助于 AIS 的途径。第三，我们在斑马鱼模型中发现基因的计划和 斑马鱼和小鼠模型中 AIS 样脊柱侧凸的功能分析将为 对人类 AIS 的原因有更机械的了解。这些假设将在三个条件下进行检验 具体目标： I. 确定 AIS 何时何地需要 Gpr126。共同祖细胞中 Gpr126 功能丧失 小鼠软骨和骨组织模型 AIS。在小鼠中使用条件遗传方法， 我们将完善这些组织中 Gpr126 的缺失如何导致脊柱侧凸的发生和进展，以及 我们将定义 Gpr126 在脊柱发育中功能的时间窗口。二.分子是什么 Gpr126在椎间盘和软骨中的功能？在这里我们将研究其机制 Gpr126 控制软骨细胞外基质的成熟和沉积以及软骨细胞的发育 椎间盘及其与小鼠脊柱侧凸发病的关系。三．分子表征 脊柱畸形的遗传学和病因学。识别对正常脊柱重要的基因和通路 在开发过程中，我们利用正向遗传学方法分离出一系列成人可行的脊柱畸形 突变斑马鱼。我们将应用大规模并行测序来识别新的脊柱畸形疾病基因 在这些现有的突变株系中。为了获得对脊柱侧凸的机械理解，我们将： a) 进行 对影响细胞外基质修饰基因的两种新型斑马鱼突变株进行功能分析； b) 在众所周知的人类 AIS 风险位点的明确斑马鱼模型中研究脊柱侧弯的病因学。 结果揭示了以前未知的正常脊柱所必需的机制和途径 发展。